Characterisation of a membrane-bound serine-specific casein kinase isolated from lactating guinea-pig mammary gland.
Serine-specific and threonine-specific casein kinase activities have been identified in a Golgi-enriched membrane fraction isolated from the lactating guinea-pig mammary gland. The serine-specific casein kinase has been purified 2000-fold by affinity chromatography on ATP-agarose. The enzyme has an estimated Mr of 100000 as determined by sucrose gradient centrifugation and phosphorylates the serine residues of dephosphorylated guinea-pig caseins A and B in a qualitatively and quantitatively identical manner to caseins A and B secreted by lactating mammary gland explants in organ culture. The enzyme also phosphorylates casein C at serine, but not threonine residues. Studies on the relative location of the enzyme within a Golgi-enriched membrane fraction show that it is an integral component of the membrane, either in the form of a transmembrane protein or exposed on the luminal side of the membrane. Although casein kinase activity is not associated with the endoplasmic reticulum, it remains to be proven whether it is truly a Golgi enzyme, since analysis of subcellular membrane components fractionated by sucrose gradient centrifugation shows that the particulate protein kinase activity of the lactating mammary gland does not cosediment with galactosyl transferase, possibly a reflection of the heterogeneous nature of mammary gland Golgi apparatus. It seems likely that the serine-specific casein kinase activity described is responsible for the phosphorylation of caseins in the lactating guinea-pig mammary gland, and that this occurs after the sequestration of processed but unphosphorylated caseins within the lumen of the endoplasmic reticulum.
D000818:Animals; D001665:Binding Sites; D047388:Casein Kinases; D055598:Chemical Phenomena; D002621:Chemistry; D005260:Female; D006056:Golgi Apparatus / Q000201:enzymology; D006168:Guinea Pigs; D007425:Intracellular Membranes / Q000201:enzymology*; D007774:Lactation; D008321:Mammary Glands, Animal / Q000201:enzymology*; D010766:Phosphorylation; D011247:Pregnancy; D011485:Protein Binding; D011494:Protein Kinases / Q000302:isolation & purification*; D012694:Serine; D013379:Substrate Specificity
D012694:Serine; D011494:Protein Kinases; D047388:Casein Kinases